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Title: Method for high-volume sequencing of nucleic acids: random and directed priming with libraries of oligonucleotides

Abstract

Random and directed priming methods for determining nucleotide sequences by enzymatic sequencing techniques, using libraries of primers of lengths 8, 9 or 10 bases, are disclosed. These methods permit direct sequencing of nucleic acids as large as 45,000 base pairs or larger without the necessity for subcloning. Individual primers are used repeatedly to prime sequence reactions in many different nucleic acid molecules. Libraries containing as few as 10,000 octamers, 14,200 nonamers, or 44,000 decamers would have the capacity to determine the sequence of almost any cosmid DNA. Random priming with a fixed set of primers from a smaller library can also be used to initiate the sequencing of individual nucleic acid molecules, with the sequence being completed by directed priming with primers from the library. In contrast to random cloning techniques, a combined random and directed priming strategy is far more efficient.

Inventors:
 [1]
  1. Stony Brook, NY
Issue Date:
Research Org.:
ASSOC UNIVERSITIES INC
OSTI Identifier:
869839
Patent Number(s):
5407799
Application Number:
08/135,317
Assignee:
Associated Universities, Inc. (Washington, DC)
DOE Contract Number:  
AC02-76CH00016
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
method; high-volume; sequencing; nucleic; acids; random; directed; priming; libraries; oligonucleotides; methods; determining; nucleotide; sequences; enzymatic; techniques; primers; lengths; 10; bases; disclosed; permit; direct; 45; 000; base; pairs; larger; necessity; subcloning; individual; repeatedly; prime; sequence; reactions; acid; molecules; containing; octamers; 14; 200; nonamers; 44; decamers; capacity; determine; cosmid; dna; fixed; set; library; initiate; completed; contrast; cloning; combined; strategy; efficient; acid molecule; nucleotide sequences; nucleotide sequence; acid molecules; nucleic acid; nucleic acids; permit direct; directed priming; base pairs; base pair; /435/

Citation Formats

Studier, F William. Method for high-volume sequencing of nucleic acids: random and directed priming with libraries of oligonucleotides. United States: N. p., 1995. Web.
Studier, F William. Method for high-volume sequencing of nucleic acids: random and directed priming with libraries of oligonucleotides. United States.
Studier, F William. Tue . "Method for high-volume sequencing of nucleic acids: random and directed priming with libraries of oligonucleotides". United States. https://www.osti.gov/servlets/purl/869839.
@article{osti_869839,
title = {Method for high-volume sequencing of nucleic acids: random and directed priming with libraries of oligonucleotides},
author = {Studier, F William},
abstractNote = {Random and directed priming methods for determining nucleotide sequences by enzymatic sequencing techniques, using libraries of primers of lengths 8, 9 or 10 bases, are disclosed. These methods permit direct sequencing of nucleic acids as large as 45,000 base pairs or larger without the necessity for subcloning. Individual primers are used repeatedly to prime sequence reactions in many different nucleic acid molecules. Libraries containing as few as 10,000 octamers, 14,200 nonamers, or 44,000 decamers would have the capacity to determine the sequence of almost any cosmid DNA. Random priming with a fixed set of primers from a smaller library can also be used to initiate the sequencing of individual nucleic acid molecules, with the sequence being completed by directed priming with primers from the library. In contrast to random cloning techniques, a combined random and directed priming strategy is far more efficient.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1995},
month = {4}
}

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